QuantideX® qPCR JAK2 Kit | Sensitive V617F Mutation Detection

The QuantideX^® qPCR JAK2 Kit offers a streamlined workflow and highly sensitive quantitative detection of the JAK2 V617F mutation. The assay design combines detection of the reference gene and target mutation in a single reaction, requiring only a single plate well for each sample. JAK2 V617F mutation testing has been the standard for the identification of myeloproliferative neoplasm (MPNs), a subset of blood cancer conditions, such as polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF). As management of these conditions has progressed, on-going testing of the JAK2 V617F mutation has gained attention. Single-gene assay kits designed to achieve higher sensitivity with process efficiency can better enable laboratories as they serve the need for extending JAK2 testing beyond identification.

Reduced Complexity

Detects the presence of the JAK2 V617F mutation relative to an endogenous control target in a single multiplex reaction
Ready-to-use design with all necessary qPCR reagents, controls, and calibrators included
Compatible with broadly installed laboratory equipment

Optimized Workflow

Multiplexed design allows for fewer run set-up steps, and more efficient use of plate-well space
Assay design harmonized with other QuantideX qPCR HemOnc portfolio kits, testing of all biomarkers on a single plate
Both peripheral blood and bone marrow samples
Accommodates a wide range of DNA input, and compatible with multiple DNA isolation methods, to fit well into your existing workflow
Automated data analysis and reporting software

Quality Results

4-point calibrator system spans assay dynamic range
Highly sensitive assay with Limit of Detection (LOD), in line with 10^-4NCCN and ELN guidelines
Individualized, sample-specific reaction LODs automatically calculated and reported by the data analysis and reporting software module

Wide Dynamic Range

Very close agreement between qPCR and reference results, with a high correlation coefficient (R2), slope close to 1.0 and intercept close to 0, illustrating the assay is linear across the full quantifiable range from the LOD/LOQ – 100% VAF.

Highly Sensitive Results

Limit of Detection/Quantitation (LoD, LoQ) of 0.0126%, using target DNA input of 165 ng, determined across 360 measurements, two lots, and three commonly available qPCR instrument platforms, with an overall detection rate of >97%.

LOD		Kit Lot
LOD		38800	38922	Overall by Platform
Platform	7500	0.0126	0.00785	0.00981
	QS5	0.0109	0.0123	0.0116
	QS7	0.00832	0.00986	0.00929
	Overall by Lot	0.0102	0.00986	0.0101

DNA Input Flexibility

DNA input range of 20–615 ng supported, with individualized, sample-specific reaction LODs automatically calculated between 0.0126% and 0.123%, dependent on DNA input.

Input (ng)	Estimated 95th Percentile Reaction LOD
200	0.0126
180	0.0126
179	0.0127
165	0.0138
100	0.0232
80	0.0292
60	0.0394
40	0.0599
20	0.123

Highly Reproducible Results

Single-site precision study across ~290 measurements, two operators, two kit lots, three run days, with high precision results for all levels, and 100% positive percent agreement (PPA).

Mean VAF	VAF CV (%)	Percent Detected
90.8	6.0	100%
74.2	6.1	100%
49.8	7.6	100%
28.7	6.3	100%
11.6	10	100%
2.57	7.5	100%
0.130	19	100%
0.0538	38	100%

QuantideX qPCR JAK2 Kit Workflow

Whole blood and bone marrow aspirate samples in K₂EDTA tubes.
Standard blood DNA extraction method prior to qPCR.
QuantideX JAK2 Analysis Software Module included with kit.
<2.5 hours including hands-on and instrument time.

Scalability with Harmonized qPCR Conditions for All QuantideX Portfolio Kits